Abstract：Objective To improve the replication level of varicella⁃zoster virus（VZV）in human diploid cell line MRC⁃5 and increase the yield of VZV vaccine by reducing the expression of interferon（IFN）related genes via optimizing the cell line MRC⁃5. Methods Interferon receptor 1（IFNAR1）silenced MRC⁃5 cell line（MRC⁃5IFNAR1⁃）was constructed by CRISPR／Cas9 gene editing technology，which was determined for the relative expression of IFNAR1 mRNA，and for those of mRNA of IFN related genes IFNβ and OAS1 after VZV infection by qRT⁃PCR to evaluate the effect of gene silencing. Gene mutation sequences were further identified by sequencing of the silenced sites. The replication of VZV in MRC⁃5 and MRC⁃5IFNAR1⁃ cell lines was compared 168 h after VZV infection by using qRT⁃PCR and plaque formation unit（PFU）assay， to evaluate the effect of MRC⁃5IFNAR1⁃cell line on VZV replication. Results The growth status of MRC⁃5IFNAR1⁃ cell line wasconsistent with that of MRC ⁃ 5 cells，and the relative expression of IFNAR1 mRNA decreased by 73%；The relative expressions of IFNβ and OAS1 mRNA in MRC⁃5IFNAR1⁃ cell line were 61% and 90% lower than those in MRC⁃ 5 cells respectively after VZV infection；In addition，168 h after VZV infection，the level of DNA replication and the titer of VZV increased by 5. 7 folds and 4 folds respectively. Conclusion The successful establishment of MRC⁃5IFNAR1⁃ cell line may be a potential scheme to increase the yield of vaccines based on human diploid cells，and provided a reference for expanding production of VZV vaccine.

OBJECTIVE: To investigate the effect of Yinlai Decoction (YD) on the microstructure of colon, and activity of D-lactic acid (DLA) and diamine oxidase (DAO) in serum of pneumonia mice model fed with high-calorie and high-protein diet (HCD). METHODS: Sixty male Kunming mice were randomly divided into 6 groups by the random number table method: normal control, pneumonia, HCD, HCD with pneumonia (HCD-P), YD (229.2 mg/mL), and dexamethasone (15.63 mg/mL) groups, with 10 in each group. HCD mice were fed with 52% milk solution by gavage. Pneumonia mice was modeled with lipopolysaccharide inhalation and was fed by gavage with either the corresponding therapeutic drugs or saline water, twice daily, for 3 days. After hematoxylin-eosin staining, the changes in the colon structure were observed under light microscopy and transmission electron microscope, respectively. Enzyme-linked immunosorbent assay was used to detect the protein levels of DLA and DAO in the serum of mice. RESULTS: The colonic mucosal structure and ultrastructure of mice in the normal control group were clear and intact. The colonic mucosal goblet cells in the pneumonia group tended to increase, and the size of the microvilli varied. In the HCD-P group, the mucosal goblet cells showed a marked increase in size with increased secretory activity. Loose mucosal epithelial connections were also observed, as shown by widened intercellular gaps with short sparse microvilli. These pathological changes of intestinal mucosa were significantly reduced in mouse models with YD treatment, while there was no significant improvement after dexamethasone treatment. The serum DLA level was significantly higher in the pneumonia, HCD, and HCD-P groups as compared with the normal control group (P<0.05). Serum DLA was significantly lower in the YD group than HCD-P group (P<0.05). Moreover, serum DLA level significantly increased in the dexamethasone group as compared with the YD group (P<0.01). There was no statistical significance in the serum level of DAO among groups (P>0.05). CONCLUSIONS YD can protect function of intestinal mucosa by improving the tissue morphology of intestinal mucosa and maintaining integrity of cell connections and microvilli structure, thereby reducing permeability of intestinal mucosa to regulate the serum levels of DLA in mice.
Mice ; Male ; Animals ; Lactic Acid/pharmacology* ; Intestinal Mucosa ; Colon/pathology* ; Dexamethasone/pharmacology* ; Diet, High-Protein ; Pneumonia/pathology*

Six dimensional syndrome differentiation theory， put forward by professor GU Xiao-hong at Beijing University of Chinese Medicine based on her theoretical teaching and clinical experience， emphasizes that the syndrome differentiation should be carried out from six dimensions including etiology， disease location， disease stage， disease condition， pathology， and disease nature， which is conducive to clinical thinking training and formation of traditional Chinese medicine （TCM）. The differentiation and treatment of Baihutang syndrome frequently seen in cold damage and warm disease still need to be explored. Guided by the six dimensional syndrome differentiation theory coupled with diverse viewpoints of cold damage and warm disease schools， this paper summarized and reinterpreted the understandings and thoughts of GU Xiao-hong and YU He， warm disease specialists of two generations. Considering the lung-stomach dysfunction caused by the internal invasion of exogenous pathogens， Baihutang syndrome was staged into Qi aspect. In this stage， exuberant pathogens and sufficient healthy Qi allowed the prevailing of internal heat and the consumption of body fluid， manifested as high fever， profuse sweating， thirst， and the pulse corresponding to interior excess and heat syndrome. This paper also pointed out that the Baihutang syndrome involved both lung and stomach， and the adoption of Baihutang contributed to preventing tu from restricting shui in the case of extreme excess of Yang brightness and protecting the kidney Yin. As revealed by the dynamic analysis of prognosis of Baihutang syndrome based on the six dimensional syndrome differentiation theory， even though the Baihutang syndrome could be present in both cold damage and warm disease， the specific disease stage， transmission and change， condition， prognosis， pathology， and medication differed. On this basis， a series of prescriptions have been modified from Baihutang， which has expanded the application scope of Baihutang and enriched its research value， thus better promoting its clinical application.

Objective To observe the effect of edaravone on endogenous neural stem cells in rats with permanent cerebral ischemia. Methods The rat model of permanent cerebral ischemia was established by electrocoagulation. Thirty SD rats were randomly divided into three groups; sham operation group, brain injury group and edaravone group. Six hours after the establishment of the model, the edaravone group was intraperitoneally injected with 1.5 g/L edaravone (10 ml/kg) once a day. The sham operation group and the cerebral ischemia group were intraperitoneally injected with saline of equal volume for 7 days. 24 hours after the last administration, BrdU positive cells, Nestin/BrdU positive cells, neuronal class III β-tubulin (Tuj1)/BrdU positive cells and glial fibrillary acidic protein (GFAP)/BrdU positive cells were observed by immunofluorescent staining, Tuj1 and GFAP protein expressions were detected by Western blotting. Results Compared with the cerebral ischemia group, the BrdU positive cells, Nestin/BrdU positive cells, Tuj1/BrdU positive cells and GFAP/BrdU positive cells increased significantly in the infraventricular area and the cortex area around the ischemia in the edaravone group (P<0.05). Compared with the cerebral ischemia group, the expression of Tuj1 and GFAP protein in the cerebral cortex of edaravone group increased ( P<0.05). Conclusion Edaravone can promote the proliferation of endogenous neural stem cells and astrocytes in the subventricular area and the cortex around ischemia, and promote the differentiation of endogenous neural stem cells into neurons.

Blood Loss, Surgical ; prevention & control ; Epilepsy ; chemically induced ; Humans ; Orthopedic Procedures ; Spine ; surgery ; Tranexamic Acid ; adverse effects

Objective:To construct the recombinant DNA E.coli ghosts (EBGs) expressing Treponema pallidum adhesin Tp0751 (pcD/Tp0751-BG) and determine its immunocompetence in immunized mice,and provide a potential novel method for syphilis vaccine developing.Methods:The recombinant eukaryotic expression plasmid pcDNA3.1(+)/Tp0751 was constructed and loaded into empty EBGs to create pcD/Tp0751-BG.The loading rate was determined accordingly.Macrophages cell line RAW264.7 was transfected with pcD/Tp0751-BG,and the expression of recombinant Tp0751(rTp0751) protein was detected by Western blot(WB).For immuno-competence in mice,the female BALB/c mice were randomly divided into 6 groups,including three control groups,A (PBS),B (EBG),C (empty pcDNA),and experimental group D(naked pcD/Tp0751),E (pcD/Tp0751-BG) and F (pcD/Tp0751-BG+rTp0751).All the mice were immunized as indicated for three times by intramuscular injection at two weeks intervals.The levels of specific IgG in sera and SIgA in genital tract lavage fluid were measured by ELISA.Levels of lymphocyte proliferation and IFN-γ secretion in spleen cells were measured by CCK-8 Cell Counting Kit and ELISA as well,respectively.Results:The loading rate of pcD/Tp0751 to EBGs was 76.1%.WB showed that the target recombinant protein pcD/Tp0751 expressed in RAW264.7 cells was active with Tp-infected rabbit sera.The titers of specific IgG and SIgA in group D,E,F gradually increased to significantly higher level as compared to the control groups (P<0. 01),which reached its peak at wk 8 after last immunization(the titers of IgG and SIgA were 1 :102 400 and 1 :12 800 in group F,respectively). Higher levels of specific IgG and SIgA were observed in groups E and F as compared to group D after first boost (P<0. 01),with groups F higher than group E after last boost(P<0. 01). At wk 8 after the last boost,the stimulation index (SI) and levels of IFN-γ in group D,E,F were all significant higher than the control groups (P<0. 01), with group E and F higher than group D (P<0. 01),and group E higher than group F (P<0. 05). Conclusion: The recombinant DNA EBGs of T. pallidum adhesin Tp0751 (pcD/ Tp0751-BG) possesses the immunocompetence to induce not only strong mucosal and systemic humoral immune response but also systemic cellular immune response in BALB/ c mice. The heterologous boost can be more efficient than homologous boost during immunization process.

OBJECTIVETo compare the clinical outcome of anterolateral minimally invasive approach versus conventional posterior approach for total hip replacement against femoral neck fractures in elderly patients.

METHODSThe retrospective study was carried out on 42 patients who suffered from displaced femoral neck fractures (19 cases of Garden type III, 23 cases of Garden type IV) treated by total hip replacement via anterolateral minimally invasive approach or conventional posterior approach by the same experienced surgeon. The average age of the patients was 78.1 years (range: 65-89 years). They were divided into anterolateral mini-invasive group (22 cases) and posterior group (20 cases). The mean time of follow-up was 13 months (range: 6-36 months). The anterolateral approach described by Hardinge goes through between anterior 1/3 and posterior 2/3 of the gluteus medius muscle, reaching the femoral neck from anterior capsule. The traditional posterior approach described by Moore (Southern incision) goes through the insertions of short external rotation muscles, reaching the femoral neck from posterior capsule. The related variables under observation were length of incision, operation time, postoperative limp, length of hospital stay and bed stay and dislolcation rate.

RESULTSThe length of the skin incision varied between 7 cm and 12 cm with the anterolateral minimally invasive technique, compared to 15-22 cm in the conventional procedure. It took less time (average 15 minutes) to complete the anterolateral minimally invasive approach (72 min+/-15 min), compared with the conventional approach (87 min+/-10 min). The average Harris hip score was 91.23+/-10.20 in anterolateral approach, 90.03+/-11.05 in the posterior approach. The average length of hospital stay for patients with the anterolateral approach was (6.4+/-2.2) days (range: 4-9 days), while that in posterior approach was (9.2+/-3.1) days (range: 6-13 days). The average length of bed stay was (3.4+/-1.1) days (range: 2-5 days) in anterolateral group and (6.2+/-2.8) days (range: 3-10 days) in posterior group. No patients in anterolateral group experienced dislocation. One (5%) hip in posterior approach had dislocation.

CONCLUSIONSAnterolateral mini-invasive approach can decrease trauma, operation time, length of hospital stay and bed stay and rehabilitation time. The stability and minimal muscular damage permit the acceleration of postoperative rehabilitation, which can subsequently reduce the perioperative risk in the treatment of femoral neck fractures in the elderly undergoing total hip replacement.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; methods ; Female ; Femoral Neck Fractures ; surgery ; Humans ; Male ; Minimally Invasive Surgical Procedures

OBJECTIVETo evaluate the effect of arthroscopic debridement for knee osteoarthritis.

METHODSThe medical records of 56 patients (56 knees) who underwent arthroscopic debridement procedures for knee osteoarthritis from 2000 to 2004 were reviewed. Among the patients, 15 patients were male and 41 patients were female,ranged in age from 39 to 76 years, with an average of (55.23 +/- 10.26) years. The duration of the disease ranged from 1 to 25 months, with an average of 5.75 months. The chief symptoms were pain, swelling and dysfunction of the knees. According to Kellgren-Lawrence classification grade, 17 patients were grade I ,39 patients were grade II. Lysholm score was used to evaluate the postoperative effects, and the statistics analysis was carried out with SPSS 13.0.

RESULTSAll the patients were followed up more than 5 years. Three patients underwent total knee replacement respectively at 5.5, 7.8 and 8.3 years after the arthroscopic debridement, the average Lysholm score of other 53 patients increased from (42.40 +/- 6.78) preoperatively to (75.53 +/- 8.23) postoperatively. There were significant difference between preoperative score and postoperative score (t = -22.62, P < 0.01).

CONCLUSIONThe good midterm follow-up results of arthroscopic debridement for knee osteoarthritis are related to the appropriate indications selections, limited debridement, good patient education, systemic rehabilitation and chondroprotective agents.

Adult ; Aged ; Arthroscopy ; Debridement ; Female ; Follow-Up Studies ; Health Education ; Humans ; Male ; Middle Aged ; Osteoarthritis, Knee ; pathology ; surgery

OBJECTIVETo investigate therapeutic effects of intra-articular injection of sodium hyaluronate, arthroscopy and total hip replacement for the treatment of different severity hip osteoarthritis.

METHODSEighty-nine patients were treated with the three methods from June 2004 to August 2008. There were 33 males and 56 females, ranging in age from 30 to 72 years, averaged 51.8 years. The disease course ranged from 3 to 360 months, with a mean of 58.7 months. All the patients had hip pain and limited movement of hip joint. The patients were treated according to ISOA score and Kellgren-Lawrance (K-L) grades of hip osteoarthritis. The preoperative and postoperative Harris scores were compared.

RESULTSAfter the operation, 83 patients were followed up, and the duration ranged from 3 to 58 months. The Harris scores results: the patients with slight hip osteoarthritis and K-L grade I got an averaged preoperative score of (70.50 +/- 4.62) and postoperative score of (91.75 +/- 4.01); the patients with slight hip osteoarthritis and K-L grade II got an averaged preoperative score of (70.15 +/- 3.79) and postoperative score of (82.18 +/- 3.92). The patients with moderate hip osteoarthritis and K-L grade III got an averaged preoperative score of (57.12 +/- 2.19) and postoperative score of (89.94 +/- 1.13) score. The patients with serious hip osteoarthritis and K-L grade IV got an averaged preoperative score of (48.63 +/- 2.21) and postoperative score of (90.76 +/- 1.14); the patients with very serious hip osteoarthritis and K-L grade IV got an averaged preoperative score of (42.67 +/- 3.17) and postoperative score of (91.97 +/- 2.24); the patients with extremely serious hip osteoarthritis and K-L grade IV got an averaged preoperative score of (36.46 +/- 2.96) and postoperative score of (82.69 +/- 1.19). There were significant differences in scores of all groups before and after treatment.

CONCLUSIONThe hip osteoarthritis should be treated according to its serious degree such as slight, moderate and severe, as well as based on its X-ray classification, which is the best method to get satisfactory results.

Adult ; Aged ; Arthroplasty, Replacement, Hip ; methods ; Arthroscopy ; Female ; Humans ; Hyaluronic Acid ; administration & dosage ; Injections, Intra-Articular ; Male ; Middle Aged ; Osteoarthritis, Hip ; therapy

OBJECTIVEEarly response to therapy is one of the most important prognostic factors in childhood acute lymphoblastic leukemia (ALL). This study aimed to assess the prognostic value of morphological assessment of bone marrow blasts during remission induction and determination of minimal residual disease (MRD) after remission induction.

METHODSFrom January 1998 to May 2003, 193 children with newly diagnosed ALL were enrolled on the ALL-XH-99 protocol. Blast cell count in the bone marrow was examined on day 19 of remission induction and by the completion of remission induction. MRD was measured with the flow cytometry. Event-free survival (EFS) was estimated by Kaplan-Meier analysis and the distributions of EFS were compared using the log-rank test. A Cox proportional hazards model was used to identify independent prognostic factors.

RESULTSThe 4-year EFS was significantly worse in patients with > or = 5% lymphoblasts in the bone marrow on day 19 as compared to those with <5% lymphoblasts on that date (42.59%+/- 14.28% vs 74.24%+/- 6.67%; p< 0.01). The 4-year EFS was significantly worse in patients with any amount of lymphoblasts in the bone marrow on the remission date as compared to that of other patients with no morphologically identifiable blasts (63.47%+/-9.23% vs 76.41%+/- 6.09%; p<0.05). The patients with MRD <0.01 had significantly better outcome than those with a level > or = 0.01% (15-month EFS:94.44%+/-5.40% vs 23.81%+/- 20.26%; p<0.01).

CONCLUSIONSEarly treatment response as assessed by morphological examination or minimal residual leukemia determination by flow cytometry has important prognostic significance, and can be performed in a resource-poor patient population.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Neoplasm, Residual ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; mortality ; pathology ; Prognosis ; Proportional Hazards Models ; Treatment Outcome